JP6754497B2 - Electrophoresis medium container - Google Patents

Description

The present invention relates to a container for an electrophoresis medium suitable for use in an electrophoresis apparatus for separating and analyzing nucleic acids, proteins and the like.

In recent years, as an electrophoresis apparatus, a capillary electrophoresis apparatus in which a capillary is filled with an electrophoresis medium such as a polymer gel or a polymer solution has been widely used.

Patent Document 1 describes that "the liquid transfer of the migration medium is performed by pressing the sealing component of the migration medium container from the outside". In Patent Document 1, the cost is reduced by using a resin molded product for the electrophoresis medium container. Further, by creating a structure that suppresses deformation on the device side in which the container is set, instead of increasing the rigidity of the electrophoresis medium container, it is possible to achieve high withstand voltage even with an inexpensive electrophoresis medium container. Further, the apparatus side is provided with a function of detecting the remaining amount of the migration medium container, and has a function of removing the internal pressure in the migration medium container after the liquid is fed. This makes it possible to control the remaining amount in the electrophoresis medium container and the amount of liquid to be sent.

International Publication No. 2016/157272

When the migration medium is sent to the capillary, the sealing component slides along the inner wall of the container syringe portion. At this time, due to the expansion of the sealing component due to the internal pressure, the contact area between the inner wall of the syringe portion and the sealing component changes, and the generated sliding resistance changes, so that there is a problem that the liquid feeding pressure is not stable during the liquid feeding. .. If the liquid feeding pressure is not stable, it becomes difficult to control the liquid feeding amount with fine resolution as in the conventional case, and the liquid feeding amount for determining that the migration medium can be filled in the capillary will vary. In particular, when the amount of the electrophoresis medium for filling varies in the direction of increasing, the running cost cannot be reduced. Further, when the electrophoresis medium leaks to the outside of the container through the gap between the inner wall of the syringe portion and the sealing component, the amount of liquid to be fed also varies. In Patent Document 1, the above-mentioned points have not been examined.

In order to solve the above problems, it is necessary to stabilize the seal sliding resistance during the transfer of the migration medium, stabilize the transfer pressure, and transfer the migration medium without leaking to the outside of the container. Therefore, an object of the present invention is to provide a migration medium container capable of stabilizing the liquid feeding pressure while ensuring the sealing property of the migration medium.

In order to solve the above problems, the migration medium container of the present invention is a migration medium container including a syringe portion for holding the migration medium and a sealing component for sealing one end of the syringe portion, and the sealing component is It has a sealing surface, a body portion, and a groove provided between the sealing surface and the body portion, and the sealing surface comes into contact with the inner wall of the syringe portion.

According to the present invention, the sealing surface comes into close contact with the inner wall of the container syringe portion, and the sealing property can be ensured.

Schematic diagram of the apparatus configuration of the present invention Top view of the device of the present invention AA sectional view of the device Detailed view of capillary array Detailed view of the electrophoresis medium container Cross section of seal parts Perspective view of seal parts Enlarged cross-sectional view of the seal surface Enlarged cross-sectional view of the seal surface Schematic diagram of the seal surface cross section when the seal parts are inserted Cross-sectional view when the seal parts are connected Perspective view of connectable seal parts Detailed view of mounting the electrophoresis medium container Schematic diagram of liquid feeding mechanism Detailed diagram of electrophoretic medium liquid feeding operation (initial state) Detailed diagram of running medium liquid transfer operation (plunger contact detection) Detailed diagram of operation of electrophoretic medium feed (injection of electrophoretic medium) Pressure graph generated during polymer feeding Pressure graph generated during polymer feeding Cross-sectional view of a sealing component provided with sealing surfaces at multiple locations

Hereinafter, examples of the present invention will be described with reference to the drawings.

FIG. 1 shows an apparatus configuration diagram of a capillary electrophoresis apparatus to which the present invention is applied. This device can be roughly divided into two units, an autosampler unit 16 at the bottom of the device and an irradiation detection / constant temperature bath unit 17 at the top of the device.

The autosampler unit 16 has a Y-axis drive body 8 mounted on the sampler base 7, a Z-axis drive body 9 mounted on the Y-axis drive body 8, and a sample tray 11 on the Z-axis drive body 9. Is installed, and the sample tray 11 can be driven on the Y-axis and the Z-axis. The user sets the electrophoresis medium container 2, the anode side buffer solution container 3, the cathode side buffer solution container 4, and the sample container 5 on the sample tray 11. The sample container 5 is set on the X-axis drive body 10 mounted on the sample tray 11, and only the sample container 5 can be driven on the X-axis on the sample tray 11. The liquid feeding mechanism 6 is also mounted on the Z-axis drive body 9. The liquid feeding mechanism 6 is arranged below the migration medium container 2.

The irradiation detection / constant temperature bath unit 17 includes a constant temperature bath unit 12 and a constant temperature bath door 14, and the inside can be kept at a constant temperature. An irradiation detection unit 15 is mounted behind the constant temperature bath unit 12 to perform detection during electrophoresis. The user sets the capillary array 1 in the constant temperature bath unit 12, performs electrophoresis while keeping the capillary array 1 at a constant temperature in the constant temperature bath unit 12, and detects it by the irradiation detection unit 15. Further, the constant temperature bath unit 12 is also equipped with an electrode 13 for dropping the voltage to GND when a high voltage for electrophoresis is applied.

As described above, the capillary array 1 is fixed to the constant temperature bath unit 12. The electrophoresis medium container 2, the anode side buffer solution container 3, the cathode side buffer solution container 4, and the sample container 5 can be driven in the YZ axis by the auto sampler unit 16, and only the sample container 5 is further driven in the X axis. Can be done. The electrophoresis medium container 2, the anode-side buffer solution container 3, the cathode-side buffer solution container 4, and the sample container 5 can be automatically connected to the fixed capillary array 1 at arbitrary positions by the movement of the autosampler unit 16. ..

FIG. 2 shows a top view of the capillary electrophoresis apparatus. The anode-side buffer solution container 3 set on the sample tray 11 includes an anode-side cleaning tank 21, an anode-side electrophoresis buffer solution tank 22, and a sample introduction buffer solution tank 23. The cathode side buffer solution container 4 includes a waste liquid tank 24, a cathode side cleaning tank 25, and a cathode side electrophoresis buffer solution tank 26. The electrophoresis medium container 2, the anode side buffer solution container 3, the cathode side buffer solution container 4, and the sample container 5 are arranged in the positional relationship as shown in the drawing. That is, the positional relationship between the anode side and the cathode side when connected to the capillary array 1 is as follows: "electrophoresis medium container 2-waste liquid tank 24", "anode side cleaning tank 21-cathode side cleaning tank 25", "anode side electricity". These are the electrophoresis buffer tank 22-cathode side electrophoresis buffer tank 26 ”and the sample introduction buffer tank 23-sample container 5.

FIG. 3 shows a cross-sectional view taken along the line AA in FIG. The electrophoresis medium container 2 is inserted and set in the guide 31 embedded in the sample tray 11. Further, in the liquid feeding mechanism 6, the plunger 32 provided in the liquid feeding mechanism 6 is arranged so as to be below the electrophoresis medium container 2.

During electrophoresis, the right side of the capillary array 1 in FIG. 3 is the cathode side, and the left side is the anode side. The auto sampler unit 16 moves to the position of "anode-side electrophoresis buffer tank 22-cathode-side electrophoresis buffer tank 26", a high voltage is applied to the cathode-side capillary array 1, and the cathode-side buffer container 4 , Electrophoresis is performed by flowing it through the anode side buffer solution container 3 through the electrode 13 to the GND.

FIG. 4 shows a detailed view of the capillary array 1. The capillary array 1 has a capillary 41 which is a glass tube having an inner diameter of about φ50 μm, and the capillary 41 has a detection unit 42. The detection unit 42 is detected by the irradiation detection unit 15. A load header 46 and a SUS pipe 47 are attached to the cathode side end of the capillary 41. The material of the load hetter 46 is preferably, for example, PBT resin, which is a resin having high insulation characteristics and a high comparative tracking index. Inside the load hetter 46, a component for conducting all the SUS pipes 47 is built in, and by applying a high voltage to the parts, a high voltage is applied to all the SUS pipes 47. Capillaries 41 are passed through the SUS pipes 47 and fixed. On the anode side, a plurality of capillaries 41 are combined into one by a capillary head 43. The capillary head 43 has a capillary head tip 45 having an acute angle and a needle shape, and a capillary head boss 44 having an outer diameter larger than that of the capillary head tip 45. The material of the capillary head 43 is preferably PEEK resin, which is a resin that is hard to chip and has rigidity and is highly stable against chemicals and analysis.

Although not shown, when the capillary array 1 is fixed to the constant temperature bath unit 12, the detection unit 42, the load header 46, and the capillary head 43 are fixed, respectively. The detection unit 42 is positioned with high accuracy so that it can be detected by the irradiation detection unit. The load header 46 is fixed so as to be conductive with a portion to which a high voltage is applied. The capillary head 43 is firmly fixed so that the tip 45 of the capillary head faces directly downward and can withstand a load. The positional relationship between the cathode side and the anode side at the time of fixing is arranged so that the plurality of capillaries 41 do not overlap each other when set in the apparatus.

FIG. 5 shows a structural diagram of the migration medium container 2. The electrophoresis medium container 2 has a syringe portion 51. The seal component 52 is inserted by pushing it from the uppermost portion (upper side of the drawing) of the syringe portion 51, and is moved to the lowermost portion of the syringe portion 51. A rubber stopper 53 is placed on the uppermost portion of the syringe portion 51, and the cap 54 is turned to seal the syringe portion 51. A film 55 is further attached on the cap 54 to seal the cap 54. The inside of the migration medium container 2 is filled with the migration medium 56, but the air 57 that enters at this time is made to collect in the upper part of the syringe portion 51. The material of the electrophoresis medium container 2 is preferably a COP resin or the like, which is a resin that can be thin-walled and has little influence on the physical properties of the material due to temperature changes.

FIG. 6A shows a structural diagram of the seal component 52 to be inserted into the syringe portion 51. The seal component 52 slides in the syringe portion 51 when the migration medium 56 is fed. As shown in FIG. 6B, the shape of the seal component 52 is a tubular shape provided with a bottom portion 59, and the cross section is U-shaped inward of the electrophoresis medium container 2. Since the bottom portion 59 is provided, it has a role of sealing the migration medium 56 inside the migration medium container 2. The outer diameter of the sealing surface 58 of the sealing component 52 is slightly wider than the inner diameter of the syringe portion 51. Therefore, when inserted, the sealing surface 58 is crushed and comes into close contact with the inner wall of the syringe portion 51. The sealing component 52 has a U-shaped cross section and has a sealing surface 58 at the uppermost portion. Therefore, when the internal pressure of the electrophoresis medium container 2 increases, the sealing surface 58 expands outward to provide the syringe portion 51. Seal. The sealing surface 58 may be a flat surface or a curved surface. The body portion 61 has a diameter smaller than the outer diameter of the sealing surface 58, and has an outer diameter that does not come into contact with the inner wall of the syringe portion 51 even when it expands outward when the internal pressure of the electrophoresis medium container 2 rises. As a result, only the sealing surface 58 comes into contact with the inner wall of the syringe portion 51. In this way, the portion that comes into contact with the inner wall of the syringe portion 51 is limited to the sealing surface 58, and the variation in the contact area between the inner wall of the syringe portion 51 and the sealing component 52 is reduced, thereby reducing the change in sliding resistance during liquid feeding. It can be suppressed and the pressure during liquid feeding can be stabilized. However, if the contact area between the sealing component 52 and the inner wall of the syringe portion 51 is reduced, the sealing property is lowered and the risk of leakage of the electrophoresis medium 56 is increased. Therefore, a groove 60 is provided between the seal surface 58 and the body portion 61. By providing the groove 60, the sealing surface 58 can be more easily expanded to the outside when the internal pressure of the electrophoresis medium container 2 rises, so that the sealing surface 58 and the inner wall of the syringe portion 51 can be more closely adhered, and the sealing property is improved. The groove 60 may have a structure in which the wall thickness of the seal component 52 is thinner than that of the seal surface 58 and the body portion 61, and may be provided inside the seal component 58. 15 (a) and 15 (b) show the measurement results of the liquid feed pressure. FIG. 15A shows a case where the body portion 61 is in contact with the inner wall of the syringe portion 51, and FIG. 15B shows a case where the body portion 61 does not contact the inner wall of the syringe portion 51. When the body portion 61 of FIG. 15A was in contact with the inner wall of the syringe portion 51, the liquid feeding pressure changed from 5MPa to 4MPa, whereas the body portion 61 of FIG. 15B was the inner wall of the syringe portion 51. It can be seen that it is stable at 4 MPa from the start to the end of the liquid feeding when it does not come into contact with.

Although the seal surface 58 is provided on the upper portion of the seal component 52, as shown in FIG. 16, the body portion seal surface 62 may be provided at a plurality of locations on the seal component 52. This makes it possible to further prevent leakage of the migration medium 56. The material of the sealing component 52 has a track record of being used for sealing the fluid of the sliding portion, etc., has a larger expansion coefficient than the material of the syringe portion 51, and is ultra-high that easily spreads outward when the internal pressure of the electrophoresis medium container 2 increases. A molecular PE resin or the like is preferable, but a rubber material may also be used.

FIG. 10 shows a detailed view of the device mounting of the electrophoresis medium container 2. When setting the electrophoresis medium container 2 in the apparatus, first, the film 55 attached to the cap 54 is peeled off. After that, it is set on the guide 31 embedded in the sample tray 11 and fixed from above so as not to rise. The guide 31 has high rigidity and plays a role of suppressing deformation of the migration medium container 2 until it comes into contact with the guide 31. When the electrophoresis medium container 2 is set in the guide 31, the gap between the outer diameter of the syringe portion 51 and the inner diameter of the guide 31 is made as small as possible. The smaller the gap, the better, but the gap is set between the outer diameter of the syringe portion 51 of the resin molded product and the inner diameter of the guide 31 which is a machined product, which is not unreasonable in processing.

FIG. 11 shows a schematic view of the liquid feeding mechanism 6. The stepping motor 102 rotates according to the number of input pulses, rotates the drive screw 91, and moves the nut 93 in a straight line. The driving method of the stepping motor 102 is, for example, 1-2 phase excitation. The nut 93 is coupled to the slider 92 and the slider 92 is coupled to the plunger 32. The position of the plunger 32 is controlled by the rotary encoder 103 integrated with the stepping motor 102. The slider 92 is connected to the linear guide 98 and moves in the axial direction of the drive screw 91. A detection plate 99 is coupled to the slider 92 and detected by the origin sensor 100 fixed to the liquid feeding mechanism base 101. The detection position of the origin sensor 100 is the origin position of the plunger 32.

A torque limiter 104 is attached to the drive screw 91 as an external load that gives rotational resistance. The torque limiter 104 has a structure in which the inner ring 96 and the outer ring 95 are coaxially fitted. The inner ring 96 of the torque limiter 104 has a hollow structure. Further, the inner ring 96 and the outer ring 95 can rotate independently. There is a constant rotational resistance between the inner ring 96 and the outer ring 95. As a method of giving rotational resistance, a magnet type that is not easily affected by wear is desirable. The drive screw 91 and the inner ring 96 rotate in synchronization with each other by passing the drive screw 91 through the hollow portion of the inner ring 96 and fitting the drive screw 91 with the inner ring 96 by a parallel pin 97 that vertically penetrates the drive screw 91. The outer ring 95 is fixed so as not to rotate by the torque limiter outer ring restraint 94 fixed to the liquid feeding mechanism base 101. This structure makes it possible to give resistance to the drive screw 91. For example, a torque limiter 104 is used, which requires a torque of 45 mN · m when the outer ring 95 is fixed and the inner ring 96 is rotated.

Subsequently, the procedure at the time of injecting the migration medium 56 will be described. The positional relationship between the plunger 32, the electrophoresis medium container 2, and the capillary head 43 at each point is shown in FIGS. 12 to 14. First, as shown in FIG. 12, the electrophoresis medium container 2 is set in the guide 31. A plunger 32 of the liquid feeding mechanism 6 is arranged directly below the electrophoresis medium container 2.

FIG. 13 shows a diagram of an initial state of a series of movements of the migration medium 56 injection operation. As described above, the seal component 52 inserted in the migration medium container 2 can slide by the movement of the plunger 32. The plunger 32 is driven by the liquid feeding mechanism 6 to bring the plunger 32 into contact with the bottom portion 59 of the seal component 52.

FIG. 14 shows a state in which the migration medium 56 is being filled into the capillary 41. After the plunger 32 is brought into contact with the bottom portion 59 of the seal component 52 in FIG. 13, the seal component 52 is further pushed up to send the electrophoresis medium 56 to the capillary 41. At this time, the inside of the migration medium container 2 becomes high pressure, and each part of the migration medium container 2 expands outward. As described above, the deformation of the syringe portion 51 of the electrophoresis medium container 2 is suppressed by the guide 31, and the deformation of the rubber stopper 53 is suppressed by the capillary head 43. Further, when the sealing component 52 is deformed by the internal pressure, the sealing component 52 is deformed so as to spread outward to be more sealed. The risk of leakage of the migration medium 56 due to the deformation of the syringe portion 51 can be reduced by making the seal component 52 have a shape and strength that have a larger expansion coefficient and are more easily deformed than the syringe portion 51 of the migration medium container 2. You can.

A multi-stage structure may be provided on the sealing surface 58. FIG. 7A shows a diagram of a multi-stage structure. The sealing surface 58 has a multi-stage structure of a concave portion 71 and a convex portion 72 along the vertical direction, and when the sealing component 52 is inserted into the syringe portion 51, the convex portion 72 is crushed and adheres as shown in FIG. Even if the migration medium 56 enters between the inner wall of the syringe portion 51, the leakage pressure can be gradually reduced, so that leakage to the lower side can be prevented. Further, by narrowing the tip of the convex portion 72, it is easily crushed by the internal pressure, and the convex portion 72 and the syringe portion can be brought into close contact with each other. The sealing surface provided with the multi-stage structure may be a flat surface as shown in FIG. 7 (a) or a curved surface as shown in FIG. 7 (b).

In the first embodiment, the number of the seal parts 52 is one, but a plurality of seal parts 52 may be connected.

A case where two seal parts 52 are connected in the vertical direction will be described with reference to FIGS. 9 (a) and 9 (b). FIG. 9A is a cross-sectional view showing a state in which two seal parts 52 (composed of an upper seal 81 and a lower seal 82) are vertically connected, and FIG. 9B is a cross-sectional view of the upper seal 81 before connection. It is a perspective view which shows the lower side seal 82. A rod-shaped insertion portion 85 is provided at the bottom of the upper seal 81, and the insertion portion 85 of the upper seal 81 is connected so as to be inserted into the lower seal 82. By this connection, the sealing surface becomes two surfaces of 83 and 84, and the sealing property of the electrophoresis medium 56 can be improved. Further, by forming the sealing surfaces of the upper seal 81 and the lower seal 82 into a multi-stage structure of the concave portion 71 and the convex portion 72 as shown in FIGS. 7 (a) and 7 (b), the upper seal 81 is tentatively formed. Even if the sealing surface 83 of the seal surface 83 is scratched or chipped and the migration medium 56 leaks downward, the lower seal 82 can prevent the migration medium 56 from leaking to the outside of the migration medium container 2. Further, by providing a distance between the seal surface 83 of the upper seal 81 and the seal surface 84 of the lower seal 82 by the insertion portion 85, more migration media can be confined when the migration medium 56 leaks. It can be kept and leakage can be prevented. In addition, when the lower seal 82 is pushed by the plunger 32, if the portion other than the center of the lower seal 82 is pushed and the lower seal 82 is hit by one side, the lower seal 82 becomes slanted, but it is inserted. The upper seal 81, which is pushed by the plunger 32 and has a distance from the bottom of the lower seal 82, is not easily affected by the balance of moments, and the sealing property is maintained. In Example 1, two seal parts are connected, but three or more seal parts may be connected. In this case, for example, a plurality of upper seals 81 are provided, and a lower seal 82 is provided at the bottom.

In addition, each above-mentioned Example may be combined. As described above, according to the present invention, the sealing surface comes into close contact with the inner wall of the container syringe portion, and the sealing property can be ensured. Further, since only the sealing surface comes into contact with the inner wall of the syringe portion, it is possible to suppress the change in the generated pressure during the transfer of the electrophoresis medium. As a result, it is possible to control the amount of liquid to be sent to the capillary of the migration medium with a fine resolution, and it is possible to reduce the running cost.

1 Capillary array 2 Electrophoresis medium container 3 Anode side buffer liquid container 4 Cathode side buffer liquid container 5 Sample container 6 Liquid transfer mechanism 7 Sampler base 8 Y drive body 9 Z drive body 10 X drive body 11 Sample tray 12 Constant temperature bath unit 13 Electrode 14 Constant temperature bath door 15 Irradiation detection unit 16 Auto sampler unit 17 Irradiation detection / constant temperature tank unit 21 Anode side cleaning tank 22 Anode side electrophoresis buffer tank 23 Anode side sample introduction buffer tank 24 Waste liquid tank 25 Cathode side cleaning tank 26 Cathode side electrophoresis buffer tank 31 Guide 32 Plunger 41 Capillary 42 Detection unit 43 Capillary head 44 Capillary head boss 45 Capillary head tip 46 Load header 47 SUS pipe 51 Syringe section 52 Sealing parts 53 Rubber stopper 54 Cap 55 Film 56 Electrophoresis medium 57 Air 58 Seal surface 59 Seal bottom 60 Groove 61 Body part 62 Body part Seal surface 71 Recessed 72 Convex part 81 Upper seal 82 Lower seal 83 Upper seal surface 84 Lower seal surface 85 Insertion part 91 Drive screw 92 Slider 93 Nut 94 Torque limiter outer ring restraint 95 Torque limiter outer ring 96 Torque limiter inner ring 97 Parallel pin 98 Linear guide 99 Detection plate 100 Origin sensor 101 Liquid transfer mechanism base 102 Stepping motor 103 Rotary encoder 104 Torque limiter

Claims (9)

The syringe part that holds the electrophoresis medium and
A seal component that seals one end of the syringe portion and
A running medium container provided with
The sealing component has an opening on the upper side and a U-shaped cross section, and the electrophoresis medium is sealed inside the migration medium container.
Further, the seal component is
Seal surface and
With the torso
It has a groove provided between the sealing surface and the body portion, and has a groove.
The sealing surface is in contact with the inner wall of the syringe portion,
When the sealing component expands due to the internal pressure of the electrophoresis medium container, the body portion does not come into contact with the inner wall.
Electrophoresis medium container. The migration medium container according to claim 1.
The outer diameter of the sealing surface is larger than the outer diameter of other parts of the sealing part and larger than the inner diameter of the syringe portion.
Electrophoresis medium container. The migration medium container according to claim 2.
The sealing surface has a multi-stage structure with irregularities along the vertical direction.
Electrophoresis medium container. The migration medium container according to claim 2.
At least one of the sealing surfaces is provided on the top of the sealing component.
Electrophoresis medium container. The migration medium container according to claim 2.
The sealing surface has a flat surface or a curved surface.
Electrophoresis medium container. The migration medium container according to claim 2.
The sealing component has a larger expansion coefficient than the syringe portion.
Electrophoresis medium container. The migration medium container according to claim 2.
A plurality of the sealing parts are connected in the vertical direction.
Electrophoresis medium container. The migration medium container according to claim 3.
The convex portion of the multi-stage structure of the sealing surface narrows toward the tip.
Electrophoresis medium container. The migration medium container according to claim 7 .
A plurality of the sealing parts are connected by a rod-shaped insertion portion.
Electrophoresis medium container.

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